贵州农业科学2025，Vol.53Issue(1)：70-75,6.DOI:10.3969/j.issn.1001-3601.2025.01.009
蜂蜜中嗜渗酵母菌TaqMan实时荧光定量PCR方法的建立
Establishment of TaqMan Real-time Fluorescent Quantitative PCR Method for Osmophilic Yeast in Honey
摘要
Abstract
[Objective]A rapid method for detecting osmophilic yeast in honey was established to provide the theoretical basis for improving the quality of honey.[Method]The specific primers were designed according to the 26S rDNA D1-D2 gene sequence of osmophilic yeast from GenBank.The obtained D1-D2 gene fragment was amplified by PCR,and the recombinant plasmid pMD-18T-JM was constructed as a positive standard.The fluorescent quantitative PCR reaction conditions were optimized to establish a TaqMan probe real-time fluorescence quantitative PCR method for osmophilic yeast,and then 160 honey samples were verified.[Result]The established TaqMan probe real-time fluorescence quantitative PCR method for osmophilic yeast in honey had good specificity and its sensitivity was up to 0.004 pg/µL.The positive rate of 160 honey samples reached 98.5%by TaqMan real-time fluorescence quantitative PCR,which was 14.4 percent points higher than that of ordinary PCR detection(84.1%of positive rate),and the sensitivity was up to 100%.[Conclusion]The established TaqMan probe real-time fluorescent quantitative PCR method for osmophilic yeast is specific,sensitive,rapid and reproducible,which is suitable for the rapid detection of osmophilic yeast in honey.关键词
蜂蜜/嗜渗酵母菌/探针/荧光定量/PCRKey words
honey/osmophilic yeast/probe/fluorescence quantification/PCR分类
农业科技引用本文复制引用
姜玲玲,罗文菊,雷露,周景瑞,艾蓉,余波..蜂蜜中嗜渗酵母菌TaqMan实时荧光定量PCR方法的建立[J].贵州农业科学,2025,53(1):70-75,6.基金项目
贵州省科技支撑计划项目"贵州黄花梨蜂蜜产品溯源和加工质量控制技术的研究"(黔科合支撑[2022]一般161) （黔科合支撑[2022]一般161）
